Our teams got a lot done over the summer weeks. Here are their recent updates.
This summer has been super busy for our team! Highlights:
Members of the team visited the Lethbridge Waste Water treatment facility in order to gain a better understanding of where our proposed technology could fit into the pre-existing infrastructure.
DNA constructs arrived and we have been busy in the lab cloning and completing protein overexpression and purification.
The team has been working on their wiki page and have made a lot of progress in the site’s design and content.
To celebrate RNA Day on August 1st, we were part of the Alberta RNA and Research Training Institute booth at the Lethbridge downtown farmers market.
Fundraising efforts are moving forward with a musical showcase!
The team will have a booth as part of the Innovation Pavilion at Whoop Up Days, where they can promote synthetic biology and our project.
Fingerprints are widely used as biometric identifiers, even in everyday life, especially with increased use in technology. This project aims at enhancing the capabilities of fingerprint analysis used in the forensic world. The core idea is to immobilize bright and colourful nanoparticles made of cadmium telluride or zinc sulfide on the fingerprint ridges. In order to label the fingerprints only, the nanoparticles are functionalized with molecules that target lysozyme, a protein that is abundant in sweat, which makes up fingerprints. We have successfully synthesized cadmium telluride nanoparticles using two different syntheses. Both procedures yielded bright particles with emission colours spanning the entire visible range of wavelengths.
Furthermore, we successfully grew cadmium sulfide shells around the nanoparticles cores which resulted in further enhancement of emission intensities. Currently, we are working on incorporating DNA aptamers that bind to lysozyme into these shells. Another labelling strategy that we are exploring utilizes molecularly imprinted polymers (MIPs) that are based around manganese doped zinc sulfide nanoparticles. The surrounding polymer is made of a silica shell that is imprinted or templated by lysozyme. These particles are expected to bind to lysozyme in fingerprints directly, without the need of an aptamer.
Our primary goal for the 2018 iGEM season is the creation and characterization of a Viral-Inspired Novel Cargo Encapsulation Toolkit (VINCEnT). Through the development of VINCEnT we will be enabling its use to address a wide range of issues such as antibiotic resistance, the negative side-effects of non-specific chemotherapeutic administration, or the potential environmental impact of using GMOs as biological control agents. To demonstrate the utility and versatility of our toolkit, we will use MS2 bacteriophage, Arc Gag, and P22 bacteriophage capsids to demonstrate small molecule encapsulation and transport, DNA transfection of cell cultures, and targeted delivery of CRIPSR machinery for genome editing, respectively. Moreover, we are currently collaborating with the UofC iGEM team to adapt our CRISPR-associated constructs for specific application within their project. We are very excited about this collaboration as it should bolster both teams’ chances of success in the iGEM Giant Jamboree in Boston.
At this time, we have our first BioBrick part in pSB1C3 (minimal Arc Gag), have run a successful test overexpression, and our first purification attempt will take place this week. The successful characterization of a minimal Arc Gag nanocage is of particular interest as it has not been experimentally purified but only predicted based on homologous sequence alignments. Thus, our team has the unique opportunity to be the first to design, test, and optimize a minimal Arc Gag purification protocol. We have made excellent progress to date and are excited to start testing encapsulation of various cargo molecules with our toolkit.
Besides work on our own project, we have also been working closely with the Lethbridge High School iGEM team. As we often share lab space with the high school team, we have taken this opportunity to provide mentorship and guidance when needed. This gives the collegiate students the ability to refine their teaching skills and aids the high school students by refining their lab techniques.
To ensure that VINCEnT is addressing the needs of potential downstream users, but is not malevolently misused or environmentally mismanaged, we have been conducting and coordinating interviews with various stakeholders including PHAC, Alberta Parks, the Lake Winnipeg Foundation, the Oldman Watershed Council, NEB, and various iGEM teams.
We also recently celebrated “RNA Day” and had team members engaging the public in downtown Lethbridge in collaboration with the Alberta RNA Research and Training Institute (ARRTI). In addition, this year we had the opportunity to present our proposed project at the RiboWest conference and participate in the iGEM meet-up in Calgary, where we received valuable feedback on our project. Moving forward, we are excited to continue development and testing of VINCEnT. We also have plans to continue public outreach by attending Whoop Up Days (in collaboration with the HS iGEM team) and by holding a Do-It-Yourself Science Night (in collaboration with Amino Labs) in Lethbridge.
UCalgary iGEM Team: Developing a safer and more effective approach to gene therapy
The 2018 iGEM Calgary team has spent the summer developing a genome engineering strategy with the intent to make gene therapy safer. Their system, which they call “Snip Equip Flip”, uses CRISPR/Cas9 to insert a “landing pad” at a targeted location on a chromosome, where the DNA-modifying protein FLP – pronounced “Flip” – can integrate therapeutic DNA. They hope that their work will highlight the value of non-viral gene therapy and encourage further development in the area.
As they’ve worked to implement a proof of concept, the team has experienced firsthand how the practice of research can deviate from textbook learning. By working through a series of experimental and logistical setbacks, the team and project have both been honed and are poised to reach even greater heights as a result of the challenges. As the end of summer approaches, the young team has redoubled their efforts to pull everything together for the iGEM competition in October.
At the North American iGEM kick-off event in July, the team met with the iGEM teams from the University of Alberta and University of Lethbridge. The team is excited to continue their collaboration with the UofL team, through which they hope to integrate their systems to demonstrate the extensibility of both.
UAlberta iGEM Team: Developing a treatment for a common parasitic disease in honeybees
Throughout the summer, Team UAlberta has been busy working on creating a biosynthetic defence system against the emerging threat of Nosema ceranae, a honey bees parasite, by engineering a porphyrin-secreting E. coli strain. N. ceranae is a species of microsporidia which causes debilitating effects on the health and normal function of afflicted honey bees. N. ceranae infections contribute to higher winter mortality rates and can be disastrous to the productivity of infected hives. The aim of our project is to biosynthetically produce protoporphyrin IX (PPIX), an intermediate in the heme synthesis pathway which is endogenous to many organisms. A chemically-synthesized derivative of PPIX has been shown to reduce the effects of N. ceranae infections which is a finding that motivated us to leverage the ubiquitous heme biosynthesis pathway to produce PPIX for use against N. ceranae infections.
As the issue of N. ceranae is prominent in our community, our team has been conducting interviews with local and commercial apiculturists, researchers, and experts across Alberta to inform our project. In fact, our multi-level consultation with these stakeholders continues to inform the direction of our experiments as their input is valuable in designing a robust treatment against N. ceranae infections. We plan to air a short documentary about our findings to make known the importance of our province’s apiculture industry and the difficult obstacles beekeepers face due to issues like N. ceranae.
We have completed the genetic design for our DNA device that will enable E. coli expressing it to overproduce PPIX. The DNA building blocks of this construct have been obtained and are currently being cloned into a single device for efficient transformation into the target E. coli strain and consequent expression of the desired porphyrin-producing phenotype. Validation of our single-vector platform is underway with encouraging results, and characterization of the system’s functionality in vitro will follow soon.
To test the effectiveness of our proposed PPIX treatment within honey bees, we also have established our own beehive to source bees for our experiments. We have instituted engineering controls and “bee-proofed” one of our labs to safely maintain and conduct tests on honey bees. Experiments for testing our PPIX treatment are currently in progress and we are confident that our experimental framework will successfully address the design problem of generating a solution which incorporates stakeholder input and provides a feasible solution to the problem of Nosema ceranae. Team UAlberta is excited to continue experimental work into Fall 2018 and present work at aGEM and the Giant Jamboree!